Solubilizate with curcumin and optionally at least one other active substance

ABSTRACT

In order to make available the health-promoting and healing properties of curcumin to the human or animal organism, also in combination with at least one further active substance, a solubilizate consists of or contains a content of curcumin equal to or smaller than 10 wt %, preferably equal to or smaller than 7.5 wt %, most preferably 6 wt %, and at least one emulsifier with an HLB value in a range below 18, preferably between 13 and 18, namely polysorbate 80 or polysorbate 20 or a mixture of polysorbate 20 and polysorbate 80, with an average diameter of the curcumin-loaded micelles ranging from 5 nm to 40 nm, preferably from 6 nm to 20 nm, most preferably from 7 nm to 10 nm, for use in particular as a dietary supplement and/or pharmaceutical drug for treating and/or preventing diseases involving inflammation, cancer and other diseases.

TECHNICAL FIELD

The disclosure relates to a solubilizate comprising curcumin andoptionally at least one further active substance. Furthermore, thedisclosure relates to a fluid containing such a solubilizate, to acapsule filled with such a solubilizate or fluid, and to a dietarysupplement and/or pharmaceutical drug containing such a solubilizate.

BACKGROUND

Curcumin is discussed as an active substance based on various potentialpharmacological properties. For example, there are indications for theantioxidant and also for the anti-inflammatory effect of curcumin aswell as for the effectiveness against viruses and bacteria as well asagainst cancer. Indications could therefore be, for example,Parkinson's, Alzheimer's, diabetes, colorectal tumors, pancreaticcancer, and liver dysfunction.

In order to be able to enter the bloodstream after oral intake, theactive substance must pass through the small intestinal blood barrier,is then metabolized in the liver and enters the hepatic vein as abioavailable fraction. The rest of the total active substance ingestedand released in the body is either degraded microbially in the intestineor eliminated with the faeces or bile.

The inventor has already created a curcumin solubilizate which hassignificantly increased bioavailability compared to native curcumin.This solubilizate is described in international patent application WO2014094921 A1. Surprisingly, it has been found in several studies thatin addition to its high bioavailability, this curcumin solubilizate inits specific formulation also has an unexpectedly greater effect on thereduction of disease symptoms which are in particular associated withinflammation or cancer.

A toxicity due to the micellization of the active substance according tothe disclosure in comparison to the native form could be ruled out onthe basis of studies with MTT assays for cell viability. Theverification of cell vitality by MTT assay is based on the reduction ofthe yellow water-soluble dye3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) intoa blue-violet water-insoluble formazan.

SUMMARY

The inventor has therefore set itself the task of providing aformulation which makes the health-promoting or curative properties ofcurcumin available for the human or animal organism, also in view of acombination with at least one further active substance. In particular,it is an object of the disclosure to provide a highest possiblebioavailability of curcumin in combination with at least one furtheractive substance.

These objects are achieved in a surprisingly simple way with asolubilizate as claimed. This solubilizate consists of or containscurcumin in a content of less than or equal to 10 wt %, preferably lessthan or equal to 7.5 wt %, most preferably 6 wt %, and at least oneemulsifier having an HLB value in the range between 13 and 18, namelypolysorbate 80 or polysorbate 20 or a mixture of polysorbate 20 andpolysorbate 80, wherein the average diameter of the curcumin-loadedmicelles is between 5 nm and 40 nm, preferably between 6 nm and 20 nm,most preferably between 7 nm and 10 nm, for use in particular as apharmaceutical drug in the treatment and/or prevention of diseasesinvolving inflammation, cancer, Alzheimer's, Parkinson's, obesity, highcholesterol, elevated blood sugar, diabetes, metabolic syndrome, and/orautoimmune diseases, multiple sclerosis (MS), for reducing visceral fat,for thermogenesis, for lowering cholesterol, in particular LDLcholesterol, and/or glucose in the blood and/or triglycerides in theblood, for improving macular pigment density, for reducing oxidativestress and/or for reducing the accumulation of fat in the hepatocytes,in particular as a pharmaceutical drug for treating and/or preventingfatty liver disease, Friedreich's ataxia, lysosomal diseases, inparticular Tay-Sachs disease, arteriosclerosis, heart diseases,arthritis.

The inventor has moreover found that the positive effect of curcumin onhealing processes or maintenance of health can surprisingly be exploitedsynergistically, at least partially, in combination with other activesubstances. In an advantageous embodiment, the disclosure provides asolubilizate consisting of or containing curcumin in a content of lessthan or equal to 10 wt %, preferably less than or equal to 8 wt %, mostpreferably 3 wt % to 7 wt %; and at least one further active substance;and at least one emulsifier having an HLB value in a range below 18,preferably between 13 and 18, in particular polysorbate 80 orpolysorbate 20 or a mixture of polysorbate 20 and polysorbate 80.

In an advantageous embodiment, the solubilizate is provided in a formthat can be administered orally, in particular in a curcumin dose in therange from 0.5 mg/kg body weight to 1 mg/kg body weight, preferably in adose of 0.81 mg/kg body weight, in particular once a day.

For the purposes of this application, the term “active substance” refersto a substance that is provided in a pharmaceutically effectiveconcentration and is preferably added for the purpose of having apharmaceutical effect. Here, the name of the respective active substanceis understood to encompass also substances that are converted in thebody into the active substance and/or into its biologically active form.

For the purposes of this application, these “active substances” includesecondary phytochemicals which are produced as chemical compounds byplants, neither in energy metabolism nor in anabolic or catabolicmetabolism. One group of secondary phytochemicals and thus activesubstances in the sense of the present application are flavonoids. Theactive substances in the sense of the present application also includenatural polyphenols such as resveratrol or the polyphenols fromlicorice, and natural phenols, in particular chalcones such asxanthohumol and also include plant extracts, i.e. substances that wereextracted from plants or parts of plants using an extractant. Theseinclude extracts from hops or from the root material of the licoriceplant. The active substance is referred to as an “extract” even if it isstill dissolved in the extractant.

The active substances in the sense of the present application alsoinclude enzymes. One example of an enzyme as an active substance in thepresent application is serrapeptase. However, the application is notlimited to this enzyme.

The extract from the resin of the frankincense tree, Boswellia serrataextract, contains several pentacyclic triterpenes which together areoften referred to as total boswellic acids (“total BAs”). The term“boswellic acids” refers to a group of chemical compounds naturallyoccurring in the resin of the frankincense trees mentioned above. Thetwo basic structures are α-boswellic acid and β-boswellic acid. Also,some derivatives of the boswellic acids are known, in particularcompounds which carry a keto group at position 11 and/or which areacetylated at position 3. Boswellic acids that are currently consideredto be significant in terms of pharmacological effects in particularinclude α-boswellic acid (αBA) and β-boswellic acid (βBA) and theirderivatives 11-keto-β-boswellic acid (KBA); CAS 17019-92-0) and3-O-acetyl-11-keto-β-boswellic acid (AKBA); CAS 67416-16-9), and3-O-acetyl-α-boswellic acid (AaBA), and 3-O-acetyl-β-boswellic acid(MBA). In particular the derivative AKBA is considered to have ananti-inflammatory effect.

In the context of the present application, the term “Boswellia”, inparticular in the term “Boswellia solubilizate” is used in the sensethat the term “Boswellia” refers to the active substances from the resinof the frankincense tree, i.e. to at least one boswellic acid and/or atleast one derivative of a boswellic acid. The term “boswellic acidsolubilizate” refers to a micellar formulation of at least one boswellicacid which may also contain at least one boswellic acid derivative.

Xanthohumol is a flavonoid naturally occurring in hops. It is aprenylated plant polyphenol which is assigned to the chalcones and hasonly been identified in hops so far. The bitter hop varieties have asignificantly higher content of xanthohumol than aroma varieties. Intests, xanthohumol was found to be effective against the emergence anddevelopment of cancer cells. In laboratory experiments, it was moreoverfound that xanthohumol is capable of protecting the nerve cells of thebrain and thus could possibly help to slow down the course of diseaseslike Alzheimer's or Parkinson's.

Licorice flavonoid oil (LFO) consisting of hydrophobe licoricepolyphenols in medium-chain triglycerides has a weight-reducing effectthat is associated with reduced body fat. Moreover, antioxidantproperties are attributed to ethanolic extracts of licorice. Licoriceflavonoid oil with a glabridin concentration of 3% has the brand name“KANEKA GLAVONOID™”. It will also be referred to as “glavonoid” below.

Resveratrol is a phytoalexin with antioxidant properties, belonging tothe polyphenols. The substance is found in grapes, for example, in theskin of red grapes even in relatively large quantities, but also inraspberries, mulberries, plums, peanuts, and Japanese Knotweed.Resveratrol can also be isolated from the grapevine itself. According tothe entry in the online encyclopedia Wikipedia, in vitro studies haveshown evidence of potential anticancer activity and beneficial effectsin diseases such as atherosclerosis, heart diseases, Alzheimer'sdisease, arthritis, and some autoimmune diseases.

Serratia peptidase or serrapeptase is a proteolytic enzyme produced bythe bacterium Serratia which lives in the intestine of the silkworm.Serrapeptase is said to have beneficial effects in relieving pain,inflammation, traumatic swelling, and excess mucus secretion by theorganism. It is said to be effective like an anti-inflammatory andanalgesic similar to acetylsalicylic acid, ibuprofen or othernon-steroidal analgesics. It is also said to induce fibrinolyticanti-inflammatory and anti-oedematous activity in the tissue. Like allenzymes, serrapeptase is sensitive to the acids produced by the stomach.Therefore, the provision in a formulation that allows gastric passage isan object of the disclosure.

The solubilizate according to the disclosure may contain one or moreboswellic acids and/or one or more boswellic acid derivatives in acontent of less than or equal to 10 wt %, preferably less than or equalto 8 wt %, most preferably 4.7 wt % to 6.6 wt %.

Due to the high proportion of Boswellia, the disclosure contemplates, inan advantageous embodiment thereof, that the solubilizate contains anextract obtained from the resin of the plant Boswellia serrata byextraction using ethyl acetate, as a source of the one or more boswellicacids and/or one or more boswellic acid derivatives, with boswellicacids being contained in a concentration of at least 85 wt % in thisextract.

The solubilizate according to the disclosure may contain xanthohumol inan amount of less than or equal to 10 wt %, preferably less than orequal to 5 wt %, most preferably 1 wt % to 3 wt %.

Due to the high proportion of xanthohumol, the disclosure contemplates,in an advantageous embodiment thereof, that the solubilizate contains anethanolic extract of hard resins from hops as a source of xanthohumol,with a xanthohumol concentration in this extract in a range between 65wt % and 95 wt %, preferably in a concentration in a range from 80% to92 wt %. In particular the products “Xantho-Flav Pure” or “Xantho-Flay”that will be discussed in more detail below can be used as a xanthohumolsource in the context of the disclosure.

The solubilizate according to the disclosure may contain a fluidcontaining licorice root extract, in particular a hydrophobic solutionof a licorice root extract, preferably glavonoid, and/or glabridin, inan amount of less than or equal to 35 wt %, preferably less than orequal to 20 wt %, most preferably from 0.3 wt % to 17 wt %.

The solubilizate according to the disclosure may contain resveratrol ina range between 1 wt % and 15 wt %, most preferably in a range between 5wt % and 10 wt %.

The solubilizate according to the disclosure may contain serrapeptase ina range of up to 3 wt %, preferably in a range between 0.1 wt % and 2 wt%, most preferably in a range between 0.18 wt % and 0.35 wt %.

The solubilizate according to the disclosure may contain coenzyme Q₁₀ ina range of up to 10 wt %, preferably in a range between 0.1 wt % and 5wt %, most preferably in a range between 0.5 wt % and 1.5 wt %.

The solubilizate according to the disclosure may contain α-liponic acidin a range of up to 10 wt %, preferably in a range between 0.1 wt % and5 wt %, most preferably in a range between 0.8 wt % and 2.5 wt %.

A solubilizate consisting of or containing curcumin and at least onefurther active substance may also be provided or employed advantageouslywithin the context of the disclosure for use as an antibiotic and/or asa pharmaceutical drug in the treatment and/or prevention of diseasesinvolving inflammation, cancer, Alzheimer's, Parkinson's, obesity, highcholesterol, elevated blood sugar, diabetes, metabolic syndrome, and/orautoimmune diseases, multiple sclerosis (MS), for reducing visceral fat,for thermogenesis, for lowering cholesterol, in particular LDLcholesterol, and/or glucose in the blood and/or triglycerides in theblood, for improving macular pigment density, for reducing oxidativestress and/or for reducing the accumulation of fat in the hepatocytes,in particular as a pharmaceutical drug for treating and/or preventingfatty liver disease, Friedreich's ataxia, lysosomal diseases, inparticular Tay-Sachs disease, arteriosclerosis, heart diseases,arthritis.

In particular, the disclosure provides the solubilizates as describedabove for use as an anti-inflammatory drug and/or as an antibioticand/or as a pharmaceutical drug with an effect against cancer,Alzheimer's, Parkinson's, obesity, high cholesterol, elevated bloodsugar, diabetes, metabolic syndrome, and/or autoimmune diseases,multiple sclerosis (MS), for lowering visceral fat, for thermogenesis,as a cholesterol-lowering pharmaceutical drug, in particular withrespect to LDL cholesterol, and/or as a pharmaceutical drug with aneffect for lowering glucose in the blood and/or triglycerides in theblood, for improving macular pigment density, for reducing oxidativestress and/or for reducing the accumulation of fat in the hepatocytes,in particular as a pharmaceutical drug with an effect against fattyliver, Friedreich's ataxia, lysosomal diseases, in particular Tay-Sachsdisease, arteriosclerosis, heart diseases, arthritis.

It has also proved to be advantageous in this context for thesolubilizate according to the disclosure that the total curcuminoidconcentration in human blood plasma measured one hour after oraladministration of 500 mg of curcumin in the form of the solubilizate asdescribed above is about 500 ng curcuminoid per mL plasma±100 ngcurcuminoid per mL plasma. The total curcuminoid concentration in humanblood plasma measured over a period of 24 hours as the area under thetotal curcumin plasma concentration vs. time curve (AUC) is in the rangefrom about 9,500 to about 10,000 nmol·h/L. Thus, the solubilizate isavailable to the body to a great extent even after oral administration.

The disclosure advantageously provides solubilizates with very goodanti-inflammatory properties. The anti-inflammatory activity measured asthe concentration of C-reactive protein (CRP) in the blood serum ofarthritic rats after a single administration of the solubilizateaccording to the disclosure in a dose of 5 mg/kg body weight of curcuminis in the range from about 2100 pg/mL to about 2500 pg/mL, and after asingle administration of the solubilizate in a dose of 10 mg/kg bodyweight of curcumin it is in the range from about 1400 pg/mL to about1800 pg/mL.

The anti-inflammatory effect measured as the concentration ofmyeloperoxidase (MPO) in the blood serum of arthritic rats after asingle administration of the solubilizate in a dose of 5 mg/kg bodyweight of curcumin is in the range from about 800 mU/mL to about 900mU/mL. These values are significantly lower than those for nativecurcumin, as will be explained in more detail below.

The anti-inflammatory activity of an inventive solubilizate comprisingcurcumin and Boswellia measured as the concentration of C-reactiveprotein (CRP) in the blood serum of arthritic rats after a singleadministration of the solubilizate in a dose of 5 mg/kg body weight ofcurcumin and 10 mg/kg body weight of boswellic acids is in a range fromabout 1200 pg/mL to about 1500 pg/mL, compared to between about 3200pg/mL and about 3500 pg/mL after administration of the same dose ofnative curcumin and Boswellia, respectively.

The anti-inflammatory effect of an inventive solubilizate comprisingcurcumin and Boswellia, measured as the concentration of myeloperoxidase(MPO) in the blood serum of arthritic rats after a single administrationof the solubilizate in a dose of 5 mg/kg body weight of curcumin and 10mg/kg body weight of boswellic acids is in a range from about 750 mU/mLto about 815 mU/mL and thus is significantly lower than the about 1150mU/mL to about 1250 mU/mL after administration of the same dose ofnative curcumin and Boswellia, respectively.

The enzyme unit (U) is a unit which has since been replaced by the katal(kat) to indicate enzymatic activity. Since the numerical values changewhen katal is used, the enzyme unit (U) continues to be used in medicineand clinical chemistry. One enzyme unit U corresponds to one micro-molesubstrate conversion per minute.

Furthermore, the disclosure offers the advantage of providing thesolubilizate according to the disclosure for use in agriculture, fishfarming and/or horticulture and/or in the field of food hygiene and/orfor use as a disinfectant and/or in the field of packaging, preferablyin the packaging of beef, poultry or fish, and/or for use as adisinfectant.

In particular for the solubilizate with curcumin as the only activesubstance it was possible to show that in terms of germ contamination offoods the treatment with the solubilizate according to the disclosureresulted in a significant reduction of the microbial load. The curcuminis effective as a photosensitizer in this case. It is applied locally toa surface infected by microbes and absorbed by the microbes there, andthen the surface is exposed to light whereby the photosensitizer isactivated, which then produces reactive oxygen species (“oxygenradicals”) in the microbes which are killed thereby.

In the food sector, for example in antimicrobial packaging, the foodsuch as meat can be sprayed with an aqueous solution of a solubilizateaccording to the disclosure and packed in transparent films made of PP,PE, PC or the like and then exposed to light, so as to keep the surfaceof already packaged food germ-free. Corresponding applications forgerm-free packaging of other items are also within the scope of thedisclosure. This includes, for example, the packaging of surgicalinstruments and similar utensils.

The advantage of the formulation as a solubilizate according to thedisclosure for this type of application is that the active substance, inparticular the curcumin, can be used in a very high aqueous dilution, sothat a color load of the treated surface can be substantially completelyexcluded. Due to the high number of very small micelles, uniform andsufficient distribution of the active substance of the solubilizate isachieved on the surface to be treated even with strong dilution. Thisallows for application of the disclosure in the field of the mostgerm-free possible packaging by way of example, in a simple and verycost-effective way.

In order to provide stable micelles of the active substances, theemulsifier content, in particular the polysorbate content, may be atleast 70 wt %, preferably in the range between 75 wt % and 95 wt %, mostpreferably in the range between 79 wt % and 88 wt % within the contextof the disclosure, depending on the further components that arecontained in the solubilizate.

Depending on the specific application, the solubilizate of thedisclosure may contain up to 20 wt %, preferably up to 15 wt % ofethanol, for example, and/or up to 25 wt %, preferably between 12 wt %and 20 wt %, most preferably up to 10 wt % of glycerol, and/oradditionally up to 10 wt %, preferably up to 7 wt % of water. With theaddition of ethanol, the content of polysorbate can be reduced, which isan advantage in view of the ADI value for polysorbate (25 mg/kg bodyweight), recommended by WHO. The content of polysorbate may also bereduced by adding glycerol.

The solubilizates of the disclosure exhibit a narrow particle sizedistribution with small mean particle size, even under the physiologicalconditions of a gastric passage; the distribution of the diameter of themicelles in a dilution of the solubilizate with distilled water in aratio of 1:500 at pH 1.1 and 37° C. ranges from about d₁₀=6 nm to aboutd₉₀=20 nm. These values were determined from a volume distribution.Details of particle size analysis of the micelles of the solubilizateswill be discussed below.

An indication of the improved bioavailability compared to compositionsof curcumin or of curcumin and at least one further active substancethat have not been micellated according to the disclosure is obtained bya measurement of turbidity of the solubilizate, which is much easieraccessible to measurement techniques. As a result of the formulationaccording to the disclosure, the turbidity of the solubilizate ispreferably less than 25 FNU, more preferably less than 3 FNU, measuredby scattered light measurement using infrared light according to thespecifications of the ISO 7027 standard at a dilution of thesolubilizate in a ratio of 1:50 or 1:500 in water under physiologicalconditions (pH 1.1 and 37° C.).

In order to facilitate oral application of the solubilizate of thedisclosure in a more simple and convenient way for the consumer orpatient, the disclosure also provides a capsule filled with asolubilizate as described above, wherein the capsule is in the form of asoft gelatin capsule or a hard gelatin capsule or a soft gelatin-freecapsule or a hard gelatin-free capsule, for example a cellulose capsule.

Moreover, in the context of the disclosure, the solubilizate accordingto the disclosure may be incorporated into other fluids, in particularliquids. The active substance-filled small micelles will be retainedwhen doing so. Thus, the disclosure also provides a fluid containing thesolubilizate as described above, wherein the fluid is selected from thegroup consisting of foods, dietary supplements, beverages, cosmetics,and pharmaceutical products. In the context of the disclosure, the fluidmay in particular comprise an aqueous dilution of the solubilizate.

The disclosure furthermore provides a method for treating and/orpreventing diseases involving inflammation, cancer, Alzheimer's,Parkinson's, obesity, high cholesterol, elevated blood sugar, diabetes,metabolic syndrome, and/or autoimmune diseases, multiple sclerosis (MS),for reducing visceral fat, for thermogenesis, for lowering cholesterol,in particular LDL cholesterol, and/or glucose in the blood and/ortriglycerides in the blood, for improving macular pigment density, forreducing oxidative stress and/or for reducing the accumulation of fat inthe hepatocytes, in particular as a pharmaceutical drug for treatingand/or preventing fatty liver disease, Friedreich's ataxia, lysosomaldiseases, in particular Tay-Sachs disease, arteriosclerosis, heartdiseases, arthritis, for improving macular pigment density, wherein themethod comprises administering to the dietary supplement consumer orpatient a solubilizate according to the disclosure, in particular in acapsule or as a fluid, in particular orally, in particular once a day.

In a preferred embodiment of the inventive method, the solubilizate isadministered to the dietary supplement consumer or patient in a curcumindose ranging from 0.5 mg/kg body weight to 1 mg/kg body weight,preferably in a dose of 0.81 mg/kg body weight.

In a preferred embodiment of the inventive method, the solubilizate isadministered to the dietary supplement consumer or patient in aBoswellia dose ranging from 1 mg/kg body weight to 2 mg/kg body weight,preferably in a dose of 1.62 mg/kg body weight.

In a preferred embodiment of the inventive method, the solubilizate isadministered to the dietary supplement consumer or patient in axanthohumol dose ranging from 0.5 mg/kg body weight to 1 mg/kg bodyweight, preferably in a dose of 0.81 mg/kg body weight.

For producing a solubilizate according to the disclosure comprisingcurcumin and at least one further active substance, it is possible toeither mix together individually prepared solubilizates, or to directlyprepare a solubilizate containing curcumin and at least one furtheractive substance.

The disclosure furthermore provides methods for producing a solubilizateas described above. If co-micellization of curcumin and at least onefurther active substance is desired, the disclosure provides thefollowing first variant of a preparation method, comprising the steps of

-   -   (a) providing polysorbate 80 and/or polysorbate 20 and/or a        mixture of polysorbate 20 and polysorbate 80;    -   (b) adding at least one further active substance, in particular        Boswellia serrata extract and/or xanthohumol;    -   (c) adding curcumin powder;    -   wherein step (a) comprises heating to a temperature in the range        from 40° C. to 62° C., preferably to a temperature in the range        from 45° C. to 57° C., most preferably to a temperature in the        range from 48° C. to 52° C.; and    -   wherein step (b) comprises keeping the temperature unchanged        compared to step a), or heating to a temperature in the range        from 60° C. to 75° C., preferably to a temperature in the range        from 61° C. to 70° C., most preferably to a temperature in the        range from 63° C. to 67° C.; and    -   wherein step (c) comprises heating to a temperature in the range        from 82° C. to 97° C., preferably to a temperature in the range        from 83° C. to 92° C., most preferably to a temperature in the        range from 85° C. to 89° C.

This preparation method allows to produce a solubilizate which is ableto form micelles loaded with curcumin and with at least one furtheractive substance, in an aqueous dilution. For this purpose, it is alsopossible to mix the at least two active substances with one another in apreparatory step under appropriately adapted temperature control, andthen to add them in combined form, as a mixture.

In particular it is possible, prior to step b), to performed a step

-   -   b1) comprising adding water at a temperature in the range from        40° C. to 62° C., preferably at a temperature in the range from        45° C. to 57° C., most preferably at a temperature in the range        from 48° C. to 52° C.

Additionally or alternatively, step b1) may comprise adding ethanol at atemperature in the range from 40° C. to 62° C., preferably at atemperature in the range from 45° C. to 57° C., most preferably at atemperature in the range from 48° C. to 52° C.

Another option for a preparation method involving co-micellization ofcurcumin and at least one further active substance is provided by thefollowing second variant of the disclosure with a method comprising thesteps of

-   -   (a) preparing a first preparation by providing polysorbate 80        and/or polysorbate 20 and/or a mixture of polysorbate 20 and        polysorbate 80 and at least a first active substance, in        particular α-lipoic acid;    -   (b) preparing a second preparation by providing water and at        least one further active substance, in particular serrapeptase;    -   (c) adding at least one further active substance, in particular        xanthohumol and/or Boswellia serrata extract and/or glavonoid        and/or resveratrol and/or coenzyme Q₁₀, and adding curcumin        powder to the second preparation from step (b);    -   (d) combining the first preparation from step (a) and the second        preparation from step c);        -   wherein the temperature is in the range from 18° C. to            22° C. during the execution of steps (a) through (d);    -   (e) heating to a temperature in the range from 80° C. to 97° C.,        preferably to a temperature in the range from 83° C. to 92° C.,        most preferably to a temperature in the range from 85° C. to 89°        C.

Step c) may further comprise adding ethanol and/or glycerol and/or MCToil and/or polysorbate 20 and/or polysorbate 80 and/or a mixture ofpolysorbate 20 and polysorbate 80.

A further option for a preparation method involving co-micellization ofcurcumin and at least one further active substance is provided by thefollowing third variant of the disclosure with a method comprising thesteps of

-   -   (a) providing polysorbate 80 and/or polysorbate 20 and/or a        mixture of polysorbate 20 and polysorbate 80 and of glycerol and        of ethanol;    -   (b) adding at least one further active substance, in particular        an ethanolic extract of hard resins from hops, in particular        Xantho-Flav Pure powder;    -   wherein step (a) comprises heating to a temperature in the range        from 40° C. to 62° C., preferably to a temperature in the range        from 45° C. to 57° C., most preferably to a temperature in the        range from 48° C. to 52° C.;    -   and wherein step (b) comprises heating to a temperature in the        range from 60° C. to 75° C., preferably to a temperature in the        range from 61° C. to 70° C., most preferably to a temperature in        the range from 63° C. to 67° C.; and    -   (c) adding curcumin powder at a temperature in the range from        70° C. to 92° C., preferably at a temperature in the range from        75° C. to 87° C., most preferably at a temperature in the range        from 78° C. to 82° C.;    -   (d) adding glavonoid under heating to a temperature in the range        from 80° C. to 97° C., preferably to a temperature in the range        from 83° C. to 92° C., most preferably to a temperature in the        range from 85° C. to 89° C.

The disclosure also relates to solubilizates which exhibit micelles inaqueous dilution loaded with curcumin alone or else with another activesubstance alone, at least immediately after their preparation.Therefore, the disclosure also provides a method for producing asolubilizate as described above by mixing a curcumin solubilizate and asolubilizate of at least one further active substance, in particular ina quantitative ratio of 1:1 of the individual solubilizates.

The disclosure will now be explained in more detail by way of exemplaryembodiments. The following components were used:

Curcumin

The product named “Turmeric Oleoresin Curcumin Powder 95%” with theproduct code EP-5001 from Green Leaf Extraction Pvt Ltd., Kerala, India,was used as the curcumin. The curcumin powder has CAS Number 458-37-7.It is a natural product obtained by solvent extraction of the rhizomesof Curcuma Longa. The curcumin content of the powder is at least 95%,according to manufacturer specifications. This curcumin content isdetermined by ASTA method 18.0.

As an alternative to the “oleoresin turmeric 95%” curcumin powder fromGreen Leaf mentioned above, it is also possible for the exemplaryembodiments described below to use, as the curcumin, 95% curcuminextract by Neelam Phyto-Extracts, Mumbai, India, or curcumin BCM-95-SGor curcumin BCM-95-CG from eurochem GmbH, Gröbenzell, Germany, orCurcuma Oleoresin 95% from Henry Lamotte OILS GmbH, Bremen, Germany, forexample.

Boswellia

In the context of the present application, the term “Boswellia” inparticular refers to an extract from the resin of the frankincenseplant. Specifically, an extract of the species Boswellia serrata wasused, which was an extract obtained by extraction with ethyl acetatefrom the resin of the plant with the botanical name Boswellia serratawith the product code “HC22519” manufactured by Frutarom Belgium N.V.,Londerzeel, Belgium. A solubilizate containing this extract is alsoreferred to as “boswellic acid solubilizate” because of its content ofboswellic acids.

Besides extracts from the resin of the frankincense plant, it is alsopossible to use boswellic acids and/or derivatives of boswellic acidsfor the purposes of the solubilizates according to the disclosure. Inparticular, the following may be considered: alpha-boswellic acid (CASnumber 471-66-9), beta-boswellic acid (CAS number 631-69-6) and theirderivatives, 3-O-acetyl-alpha-boswellic acid (CAS number 89913-60-0),3-O-acetyl-beta-boswellic acid (CAS number 5968-70-7),11-keto-beta-boswellic acid (KBA, CAS number 17019-92-0), and3-O-acetyl-11-keto-beta-boswellic acid (AKBA, CAS number 67416-61-9).

Xanthohumol

The products “Xantho-Flav” or “Xantho-Flav Pure” of the brand“Hopsteiner” by Simon H. Steiner, Hopfen, GmbH, Mainburg, Germany wereused as the xanthohumol source. Both are natural products produced fromhops. The active substance is the hop polyphenol xanthohumol. This is ayellow colored powder with a xanthohumol content between 65% and 85% in“Xantho-Flav” and at least 85% in “Xantho-Flav Pure”, according tomanufacturer specifications.

The concentrations of xanthohumol and isoxanthohumol in “Xantho-FlavPure” are quantified by the manufacturer according to UVspectrophotometric analysis or HPLC EBC 7.8 using external calibrationstandard pure XN (370 nm) or IX (290 nm). “Xantho-Flav Pure” containsthe prenylated flavonoid xanthohumol in a very high concentration. Forthe exemplary embodiments in the context of the present application,“Xantho-Flav Pure” of batch number 9432 was used.

Glavonoid/Glabridin

“Glavonoid” is the product name for a composition of Kaneka Corporation,Osaka, Japan, which contains glabridin as an active substance. Glabridinis a flavonoid of the licorice plant (Glycyrrhiza glabra). The product“Kaneka Glavonoid” contains 30% of licorice extract and 70% of edibleoil, according to the manufacturer. “Kaneka Glavonoid” is standardizedto 3% glabridin, according to the manufacturer, which is the maincomponent of the polyphenols of the licorice plant. The CAS number ofglabridin is 59870-68-7.

Resveratrol

The product “eveResveratrol” was used as resveratrol. This istransresveratrol obtained by fermentation. The product is provided as awhitish powder without additives. The content of transresveratrol is atleast 98 wt %, the rest is water. The product has CAS number 501-36-0and product code RSXOL 9800.

Serrapeptase

The product named Serratiopeptidase from Shaanxi Pioneer Biotech Co.Ltd. with batch number PBD 20170708 was used as serrapeptase. This is agreyish white to light brown powder.

Coenzyme Q₁₀

Coenzyme Q₁₀ was purchased from Xiamen Kingdomway Group Company. It wasproduced by microbial fermentation and contains less than 0.5% of thecis isomer, according to the manufacturer's specifications.

α-Lipoic acid (alpha-liponic acid)

Alpha-lipoic acid was purchased from Jiangsu Tohope Pharmaceutical Co.Ltd., China.

Polysorbate 80

The source of polysorbate 80 was the material “TEGO SMO 80 V FOOD” withthe specification code “K04 EU-FOOD” from Evonik Nutrition & Care GmbH,Essen, Germany. The product complies with the EU requirements for foodadditive E 433. As an alternative to the TEGO SMO 80 V from Evonikmentioned above, it is also possible to use TEGO SMO 80 V from InCoPAGmbh, Illertissen, Germany, or Crillet 4/Tween 80-LQ-(SG) from CRODAGmbH, Nettetal, Germany, or Lamesorb SMO 20 and Kotilen-O/1 VL fromUnivar or from Kolb Distributions AG, Hedingen, Switzerland, as thepolysorbate 80 in the exemplary embodiments described below.

Polysorbate 20

The source of polysorbate 20 was the material “TEGO SML 20 V FOOD” withthe specification code “K09 EU-FOOD” from Evonik Nutrition & Care GmbH,Essen, Germany. The product complies with the EU requirements for foodadditive E 432. As an alternative to the TEGO SML 20 from Evonikmentioned above, it is also possible to use Crillet 1/Tween 20-LQ-(SG)from CRODA GmbH, Nettetal, Germany, as the polysorbate 20 within thecontext of the disclosure.

Ethanol

In the context of the present application, ethanol was purchased fromBerkel Pfalzische Spritfabrik GmbH & Co. KG. According to thespecification for “undenatured neutral alcohol 1411U taxed”, the contentof ethanol of this product is about 92.6 to 95.2 wt %.

Glycerol

The product used as glycerol in the context of the present applicationwas “Glycamed 99.7%” from Glaconchemie GmbH, Merseburg, Germany. Theglycerol content of this product is at least 99.5%, according tomanufacturer specifications.

Medium-Chain Triglycerides

Medium-chain triglycerides (MCTs) are triglycerides that containmedium-chain fatty acids. Medium-chain fatty acids include caproic acid,caprylic acid, capric acid and lauric acid. These are saturated fattyacids which naturally occur in tropical vegetable fats such as coconutoil and palm kernel oil. To a small extent they are also contained inmilk fat. There is no pure MCT oil in nature, however, pure MCT oils canbe obtained synthetically. Individual MCTs or a mixture of differentMCTs can be used as medium-chain triglycerides within the scope of thedisclosure. Medium-chain triglycerides were used in the form of MCT oilDelios VK Kosher, manufactured by Cognis GmbH, Monheim, Germany, or inthe form of MCT oil (70/30) Rofetan GTCC 70/30 manufactured by DHWDeutsche Hydrierwerke Rodleben GmbH, Dessau-RoBlau, Germany, CAS number73-398-61-5.

Furthermore, medium-chain triglycerides can be used in the form of theproduct ROFETAN DTCC 70/30 (Ph. Eur.). This is a caprylic/capric acidtriglyceride with CAS number 73398-61-5. The product corresponds to themonograph “medium-chain triglycerides” of the European Pharmacopoeiavalid at the filing date. Manufacturers are Ecogreen Oleochemicals DHW,Deutsche Hydrierwerke GmbH, Rodleben, Germany.

Mixed Tocopherol

The 70% mixed tocopherol in vegetable oil Vitapherole T-70 Non GMO,manufactured by VitaeNaturals can be used as a mixed tocopherol (E306,CAS numbers 59-02-9, 16698-35-4, 54-28-4, and 119-13-1), for example.

If water is added in the preparation of a solubilizate, distilled wateris used.

Prof. Dr. Ing. M. T. Khayyal from the University of Cairo, Faculty ofPharmacy at the Institute for Pharmacology, performed studies on theanti-inflammatory effect of curcumin and combinations of curcumin withBoswellia or xanthohumol, in each case in the native form and in thesolubilized form according to the disclosure.

Anti-inflammatory markers and antioxidant capacity were determined.Female Wistar rats with a body weight between 150 and 200 g were exposedto adjuvant induced arthritis according to Pearson et al. (1956). At day0, the animals were administered 0.1 ml of Freund's Adjuvant (FCA) inthe right hind paw, by subplantar injection. The animals were randomlydivided into 12 groups of 8 animals each.

Group 1 was the control group.

Group 2 received diclofenac as a reference drug in a dose of 3 mg/kgbody weight.

Group 3 received native curcumin in a dose of 5 mg/kg body weight.

Group 4 received curcumin solubilized according to the disclosure in adose of 5 mg/kg body weight.

Group 5 received native curcumin in a dose of 10 mg/kg body weight, and

Group 6 received curcumin solubilized in the same dose.

Group 7 received native xanthohumol in a dose of 5 mg/kg body weight,and

Group 8 received solubilized xanthohumol in the same dose.

Group 9 received a mixture of native curcumin in a dose of 5 mg/kg bodyweight and native Boswellia extract in a dose of 10 mg/kg body weight.

Group 10 received a mixture of solubilized curcumin and solubilizedBoswellia in the same respective dose.

Group 11 received a mixture of native curcumin in a dose of 5 mg/kg bodyweight and native xanthohumol in a dose of 5 mg/kg body weight, and

Group 12 received a mixture of solubilized curcumin and solubilizedxanthohumol in the same respective dose.

All extracts or solubilizates were administered orally once daily fromday 0 to day 21 following the vaccination with the adjuvant. After day21, the animals were killed and serum samples were prepared and storedat −80° C. Measurements were made of myeloperoxidase (MPO), C-reactiveprotein (CRP), total antioxidant capacity (TAC), and thiobarbituraticacid reactive substances (TBARS).

BRIEF DESCRIPTION OF THE DRAWINGS

The results will now be explained with reference to the accompanyingfigures, wherein:

FIG. 1a illustrates the effect of curcumin in native and in solubilizedform and of diclofenac on the serum CRP level (pg/L).

FIG. 1b illustrates the effect of curcumin (5 mg/mL) in native and insolubilized form when administered together with either Boswellia orxanthohumol on the serum CRP level (pg/L), compared to diclofenac.

FIG. 2 illustrates the effect of curcumin in native and in solubilizedform and of diclofenac on the serum MPO level (mU/mL).

FIG. 3 illustrates the effect of curcumin in native and in solubilizedform when administered together with either Boswellia or xanthohumol onthe serum MPO level (mU/mL), compared to diclofenac.

DETAILED DESCRIPTION

First, the effects on C-reactive protein (CRP) were studied. C-reactiveprotein is a specific marker for anti-inflammatory activity. Both thenative and the solubilized forms of curcumin inhibited rat serum CRPlevels in a dose-dependent manner, but the solubilized form was twice aseffective as the native form and significantly more effective thandiclofenac at the chosen dose (FIG. 1a ).

When Boswellia is administered together with curcumin in their nativeforms, the inhibitory effect on serum CRP does not change significantly(FIG. 1b ). However, when curcumin and Boswellia are administered insolubilized form together and at an appropriate dose, theanti-inflammatory effect is enhanced. In this regard, Boswellia issuperior to xanthohumol in potentiating the effect of curcumin at theconsidered doses.

Myeloperoxidase (MPO) in plasma plays a central role as apro-inflammatory mediator in rheumatoid arthritis and is an indicator ofthe invasion of neutrophil granulocytes into the affected tissue. Itsconcentration is elevated in patients with rheumatoid arthritis andcauses oxidative stress. In the studies, the MPO concentration waseffectively reduced by solubilized curcumin, at both dosages consideredin the same way and not significantly different from diclofenac.However, native curcumin had no effect on the MPO levels (FIG. 2). Theadministration of Boswellia together with curcumin, both in the nativeand the solubilized forms, did not improve the effect of curcumin inreducing MPO levels.

Oxidative stress is one of the major factors contributing to jointdestruction in rheumatoid arthritis (RA). An increase in the productionof so-called “reactive oxygen species (ROS)” leads to a reduced supplyof endogenous antioxidants and ultimately results in the destruction ofcells. The neutrophil granulocytes released in the rheumatoid jointproduce free oxygen radicals which cause increased formation of lipidperoxides manifesting in an increase in serum TBARS. Therefore, anincrease in antioxidant status represented by an increase in TAC can beused as an indication of protection against the development ofdegenerative inflammatory processes. There is an inverse relationshipbetween the levels of TAC and TBARS, a high level of antioxidantcapacity TAC corresponds to a low TBARS concentration.

The studies that were performed showed that, at both dosages considered,the native form of curcumin had no significant effect on the levels ofTBARS or TAC. Solubilized curcumin according to the disclosure reducedthe TBARS level at both selected dosages and increased TAC, with hardlyany differences to the effect of diclofenac.

These data are summarized in the table below. The table contains data onthe effect of curcumin and Boswellia in native and in solubilized form,administered either alone or in combination with diclofenac in a dose of3 mg per kg body weight once daily for 21 days, on the antioxidantcapacity TAC and the thiobarbituric acid reactive substances TBARS inthe serum of arthritic rats (n=8). Indicated are mean values±standarderror of the mean (SEM).

TAC TBARS Group (nmol/microliter) (nmol/L) Arthritic control group 57.26± 3.36 13.10 ± 0.39  Diclofenac (3 mg/kg) 82.08 ± 2.96 7.93 ± 0.84Native curcumin (5 mg/kg) 60.31 ± 3.25 11.64 ± 0.39  Solubilizedcurcumin (5 mg/kg) 77.01 ± 0.73 5.97 ± 0.47 Native curcumin (10 mg/kg)68.41 ± 1.09 13.18 ± 0.46  Solubilized curcumin 87.15 ± 5.27 6.82 ± 0.56(10 mg/kg) Native curcumin (5 mg/kg) + 64.56 ± 1.45 12.08 ± 0.52 Boswellia (10 mg/kg) Solubilized curcumin (5 mg/kg) + 76.94 ± 2.17 6.81± 0.19 Boswellia (10 mg/kg)

According to the results presented in the table, the administration ofBoswellia together with curcumin, each in native form, did not show anysignificant effect for reducing the oxidative stress. In solubilizedform, however, these combinations were as effective as diclofenac inreducing TBARS and increasing TAC in arthritic rat's serum.

First studies on the use of the curcumin solubilizate according to thepresent disclosure in combating cancer cells of the lines (MCF-7) and(T47D) for breast cancer, (Hepg-2) for liver cancer, (HCT-116) for coloncancer, and (PC3) for prostate cancer show very good results in terms ofachieving the lowest possible proportion of cells that survive thetreatment. The curcumin solubilizate permitted a reduction to a“surviving fraction” in the range from about 15% to about 25%.

The particle size analyzes of the micelles in aqueous dilutions ofsolubilizates according to the disclosure were measured according to theprinciple of dynamic light scattering using laser light of 780 nmwavelength, unless stated otherwise. The particle size measurements wereperformed using the ParticleMetrix NANO-flex backscatter particleanalyzer. The measuring principle is based on dynamic light scattering(DLS) in a 180° heterodyne backscattering setup.

For the experimental determination of turbidity of the solubilizatesaccording to the disclosure, the turbidity meters are calibrated with astandard suspension. Thus, instead of measured light intensity, theconcentration of the calibration suspension is indicated. So, when anyarbitrary suspension is measured, the indication means that therespective liquid causes the same light scattering as the standardsuspension at the indicated concentration. The internationally definedturbidity standard is formazine. The most common units include theindication FNU, i.e. Formazin Nephelometric Units. This is the unit usedin water treatment, for example, for measuring at 90° in compliance withthe requirements of the ISO 7072 standard.

For preparing a solubilizate according to the disclosure including theactive substances curcumin and at least one further active substance itis possible to either mix individually prepared solubilizates with oneanother or to directly prepare a solubilizate containing curcumin and atleast one further active substance or several further active substances.

Curcumin Solubilizates

By way of example, a 7% curcumin solubilizate is prepared. To this end,

925 g polysorbate 80 and 75 g curcumin powder 95% (=71.2 g of curcumin)are used. The polysorbate 80 is heated to 48 to 52° C. The curcuminpowder is added to the polysorbate under stirring, while further heatingto a temperature in the range from 95 to 97° C. The powder is added atan appropriate rate so as to be evenly drawn into the emulsifier duringstirring. After cooling to a temperature below a maximum of 60° C., thecurcumin solubilizate is bottled. This solubilizate was used for thepreparation of a curcumin and Boswellia solubilizate.

At a 1:500 dilution in water at pH 1.1 and a temperature of 37° C., the7% curcumin solubilizate exhibits an average turbidity of 0.9 FNU.

However, it should be noted that the curcumin content can be furtherincreased without having to accept adverse consequences, for example interms of stability of the micelles. A composition consisting of 100 g of95% curcumin powder and 900 g of polysorbate 80 results in a stableproduct just like a composition consisting of 120 g of 95% curcuminpowder and 880 g of polysorbate 80, or 70 g of 95% curcumin powder and930 g of polysorbate 80.

Moreover, the polysorbate 80 may be entirely or partially replaced bypolysorbate 20. For example, for preparing a curcumin solubilizate withpolysorbate 20 alone, 894 g of polysorbate 20 and 106 g of 95 curcuminpowder can be used. The polysorbate 20 is heated to between about 63° C.and about 67° C. While stirring, the curcumin powder is slowly added tothe polysorbate 20. While adding the curcumin powder, heating iscontinued to between about 83° C. and about 87° C. The resultingsolubilizate is slowly cooled to below about 45° C. and is then readyfor being bottled.

Otherwise, the preparation of these variants corresponds to thatdescribed above. Solubilizates of up to about 11% can be produced inthis way.

1.5% Serrapeptase Solubilizate

The following is used:

15 g serrapeptase: serratiopeptidase 20,000 U/mg = 300,000,000 U, 15 gwater, 16.5 g MCT oil, 953.5 g polysorbate 80.

At a temperature in the range between 18 and 22° C., water is mixed withserrapeptase, and the mixture is homogenized. This means that theserrapeptase is distributed as evenly as possible in the water. Thiscreates the conditions for the serrapeptase to be largely completelydissolved in the water. While heating to a temperature in the range from83 to 87° C., MCT oil is incorporated into the water-serrapeptasemixture under constant stirring. The stirring is performed intenselyenough for the serrapeptase to dissolve evenly in the water. Atunchanged temperature, polysorbate 80 is added under stirring and ishomogenized. The stirring is performed intensely enough for thepolysorbate 80 to be evenly distributed. The product is cooled to atemperature below 60° C. and bottled. It is then stored in the dark atnot more than 25° C.

300,000 U/g corresponds to 15 mg/g of 1.5% serrapeptase in enzymaticunits. At a dilution in water of 1:50, the turbidity of thissolubilizate was determined under physiological conditions at pH 1.1 and37° C. The resulting value was 1.8 FNU.

For a particle size analysis of the serrapeptase solubilizate, thissolubilizate was first diluted with distilled water in a ratio of 1:500and heated to 37° C. under constant stirring using a magnetic stirrerand a hotplate. Subsequently, the pH was adjusted to 1.1 using 32%hydrochloric acid. The samples were then measured immediately. Theresults are summarized in the table below, for which the data of twomeasurements were averaged.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 9.23 10.4712.23 13.56 Volume distribution 9.10 10.18 11.82 13.12

10% Resveratrol Solubilizate

The following is used:

100 g resveratrol, 45 g MCT oil, 600 g polysorbate 80, 180 g polysorbate20, and 75 g mixed tocopherol.

At a temperature in the range between 18 and 22° C., the polysorbates,the mixed tocopherol and the MCT oil are mixed and homogenized whilestirring sufficiently so that the components are evenly distributed. Theresveratrol is added to the mixture or solution of polysorbate, MCT oiland mixed tocopherol while heating to a temperature in the range from 83to 87° C., while stirring sufficiently so that the resveratrol is evenlydrawn into the emulsifier-containing preparation. Once a homogeneous andtransparent product is obtained, it is cooled to a temperature below 30°C. and bottled. The product is yellowish, transparent and viscous and isstored in the dark at a temperature of 25° C.

If, for therapeutic purposes, the administration of 3,500 mg of thenative form of the resveratrol raw material is taken as a basis, thefollowing calculation applies to the equivalent amount of solubilizate:

3 capsule fillings, each with 675 mg of solubilizate, correspond to anamount of 2,025 mg per day or 200 mg of the active substance resveratroland 100 mg of mixed tocopherol. With a DV factor of 1:17 the followingapplies: 200 mg×17=3,400 mg of resveratrol pure substance.

Turbidity of this solubilizate was also determined under physiologicalconditions (pH 1.1; 37° C.) at a dilution in water of 1:50. Theresulting value was 16.1 FNU.

5% Coenzyme Q₁₀ Solubilizate

The following is used:

57.5 g coenzyme Q₁₀, 160 g MCT oil, and 782.5 g polysorbate 80

The polysorbate is heated to a temperature in the range between 83° C.and 87° C. Then the coenzyme Q₁₀ powder is incorporated under stirring.Stirring is performed intensely enough so that the components are evenlydistributed. While maintaining the temperature or reheating to atemperature in the range from 83 to 87° C., the coenzyme Q₁₀ isdissolved in the polysorbate 80. Then, the MCT oil is incorporated whilemaintaining the temperature. Once a homogeneous and transparent productis obtained, it is cooled to a temperature below 60° C. and bottled. Theproduct is orange-red, transparent and partially solid at roomtemperature. It is stored in the dark at a temperature of 25° C.

The measurement of turbidity under physiological conditions (pH 1.1; 37°C.) at a dilution in water of 1:50 gave a value of 11.9 FNU as the meanvalue of three measurements (11.4 FNU; 10.5 FNU; 13.9 FNU).

A particle size analysis of a sample of 20 microliters in a dilutionratio of 1:50 in a solution of table salt (50 mmol/L) and sodium azide(200 mg/L) by Wyatt Technology Europe GmbH in the Eclipse program, i.e.by field flow fractionation, gave a peak with a radius of 16.5 nm forthe micelles of this solubilizate. Accordingly, the diameter of themicelles is 33 nm.

10% α-Liponic Acid Solubilizate

The following is used:

896.7 g polysorbate 80, 103.3 g α-lipoic acid.

The polysorbate 80 is heated to between 28 and 32° C. Under stirring,the α-lipoic acid powder is added to the polysorbate and incorporated.The powder is added at such a rate that it is evenly drawn into theemulsifier during stirring. This is followed by heating up to atemperature in the range between 83° C. and 87° C. Once a homogeneousand transparent product is obtained, it is cooled to a temperature below60° C. and bottled. The product is yellow and viscous and is stored inthe dark at a temperature of 25° C.

The product can also be produced using polysorbate 20 or a mixture ofpolysorbate 80 and polysorbate 20.

At a dilution ration of 1:50 in water at pH 1.1 and a temperature of 37°C., the solubilizate exhibits an averaged turbidity of 2.9 FNU.

A particle size analysis of a sample of 20 microliters in a dilutionratio of 1:50 in a solution of table salt (50 mmol/L) and sodium azide(200 mg/L) by Wyatt Technology Europe GmbH in the Eclipse program, i.e.by field flow fractionation, gave a peak with a radius below 10 nm forthe micelles of this solubilizate. Accordingly, the diameter of themicelles is not more than 20 nm.

10% Xantho-Flav Pure Solubilizate (Corresp. To 9.2% Xanthohumol) withEthanol

For this variant of a xanthohumol solubilizate according to thedisclosure, the following was used:

100 g Xantho-Flav Pure ({circumflex over (=)}92 g of xanthohumol), 150 gethanol (96%) of neutral alcohol grade 1411U, and 750 g polysorbate 80.

First, the Xantho-Flav Pure powder is dissolved in ethanol while beingheated to a temperature in the range between 48 and 52° C. A homogeneoussolution is created. Polysorbate 80 is then added into the solution ofXantho-Flav Pure in ethanol while heating to between 83 and 87° C. Theadding is done at a rate such that the two fluids homogenize well understirring. The resulting solubilizate is cooled to below 60° C. and isbottled and stored in the dark and cool, i.e. at temperatures below 25°C.

15% Glavonoid Solubilizate (=0.45% Glabridin) with Glycerol

The following was used:

150 g glavoniod (=4.5 g glabridin), 100 g 99% glycerol, 750 gpolysorbate 80.

First, glycerol and glavonoid were mixed and homogenized at atemperature in the range from 18 to 22° C. While heating up to 83-87°C., polysorbate 80 was added to the fluid consisting of glycerol andglavonoid, under stirring. The stirring was performed intensely enoughso that a homogeneous solubilizate was obtained, which was allowed tocool to a maximum of 30° C. and bottled and then stored in the dark at atemperature below 25° C.

The solubilizates described above can be used to prepare thesolubilizate according to the disclosure comprising curcumin and atleast one further active substance by mixing. This will be describedbelow with reference to exemplary embodiments 3, 5, 6, and 7.

Exemplary Embodiment 1 Solubilizate of 5.4% Curcumin/6.6% Boswellic Acid

This exemplary embodiment of the solubilizate according to thedisclosure was prepared directly. The active substances wereco-micellized. To this end, the following was used here:

82 g 80% Boswellia serrata extract (=65.6 g boswellic acid), 57 g 95%curcumin powder (=54.1 g of curcumin), 70 g water, 350 g polysorbate 20,441 g polysorbate 80.

While heating to a temperature in the range from 48 to 52° C.,polysorbate 20 and polysorbate 80 are homogenized with each other andthereby dissolved in each other under stirring. While maintaining thetemperature, the emulsifier mixture is mixed with the water whilestirring intensely enough so that the water and the ethanol aredissolved evenly in the emulsifier solution. At unchanged temperature,the Boswellia serrata extract is incorporated into the water-dilutedemulsifier under stirring. The Boswellia serrata extract is added at arate slow enough to be evenly drawn into the dilute emulsifier solutionunder stirring. Subsequently, the temperature is increased to a rangebetween 63° C. and 67° C. under vigorous stirring. The curcumin powderis incorporated under stirring. The temperature is further increased toa value in the range between 85° C. and 89° C. while stirring intenselyenough for the curcumin to be evenly distributed in the preparation andhomogenized.

At a dilution ration of 1:500 in water at pH 1.1 and a temperature of37° C., the solubilizate exhibits an averaged turbidity of 1.9 FNU.

In the context of the present application, a verification about whetherthe homogenization of the components to form a solubilizate according tothe disclosure has been sufficiently completed in the preparation of anysolubilizates is obtained by measurements of the clarity of the product,which indicates complete micellization, using a laser beam. Such a laserbeam measurement may be performed, for example, by illuminating thesample using a commercially available laser pointer, in particular witha wavelength in the range between 650 nm and 1700 nm (spectral colorred), and subsequent visual inspection of the illuminated or irradiatedsolubilizate. The verification is not achieved by sampling and thusoutside the reaction vessel, but in the reaction vessel. The laser beamis directed through a sight glass which is located on the front of thereaction vessel, perpendicularly to the reaction vessel. If merely apoint of light appears on the rear inner surface of the reaction vessel,completely free of scattering, the resulting particle structures in thereaction vessel are smaller than the wavelength of the visible light,which is thus a visual confirmation that the process of micellizationhas been completed.

In the context of the disclosure, the contents of curcumin and Boswelliaextract in the individual solubilizates may also be adjusted so as to besignificantly higher than in the example shown, depending on theapplication case.

Exemplary Embodiment 2

Solubilizate of 3.3% Curcumin/3.6% Boswellic Acid with 1.8% Xanthohumol

The following is used:

45 g 80% Boswellia serrata extract (36 g boswellic acid), 35 g 95%curcumin powder (33.25 g of curcumin), 23 g Xantho-Flav with at least80% xanthohumol (18.4 g xanthohumol), 60 g water, 50 g ethanol (96%)neutral alcohol, grade 1411U, 350 g polysorbate 20, 437 g polysorbate80.

While heating to a temperature in the range from 48 to 52° C.,polysorbate 20 and polysorbate 80 are homogenized with each other whilebeing dissolved in each other, under stirring. While maintaining thetemperature, the emulsifier mixture is mixed with the water and ethanol.Stirring is performed intensely enough so that the water and the ethanolare dissolved evenly in the emulsifier solution. At unchangedtemperature, the Boswellia serrata extract and the xanthohumol areincorporated into the water-diluted emulsifier mixture while stirring.The adding occurs at a rate slow enough so that the Boswellia serrataextract and the xanthohumol are evenly drawn into the dilute emulsifiersolution, under stirring. Subsequently, the temperature is increased toa range between 63° C. and 67° C. under vigorous stirring. The curcuminpowder is incorporated while stirring. The temperature is furtherincreased to a value in the range between 85° C. and 89° C. whilestirring intensely enough so that the curcumin is evenly distributed inthe preparation and homogenized.

This is followed by cooling to a temperature of less than or equal to45° C. The dark yellow, viscous preparation comprising a solubilizate ofcurcumin and boswellic acid and xanthohumol is then bottled and storedin the dark and cool, i.e. below 25° C.

At a dilution ratio of 1:500 in water at pH 1.1 and a temperature of 37°C., the solubilizate exhibits an averaged turbidity of 1.9 FNU.

For particle size analysis of a solubilizate according to thedisclosure, unless stated otherwise, this solubilizate was first dilutedwith distilled water in a ratio of 1:500 and brought to 37° C. underconstant stirring with a magnetic stirrer and using a hot plate.Subsequently, the pH was adjusted to 1.1 using 32% hydrochloric acid.The samples were then measured immediately. The results are summarizedin the table below.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 10.18 15.70533.0 3080 Volume distribution 7.90 10.96 15.21 20.37

Exemplary Embodiment 3 Solubilizate of 1.5% Curcumin/3% BoswellicAcid/2% Xanthohumol/0.35% Serrapeptase

The following is used according to the formulations described above:

250 g 7% curcumin solubilizate, 250 g 12% Bowellia solubilizate, 250 g9.2% xanthohumol solubilizate, and 250 g 1.5% serrapeptase solubilizate.

All four solubilizates can be heated to a temperature in the range from50° C. to 60° C. to lower viscosity and thus enhancing flowability.Subsequently, they are mixed together by stirring. As soon as ahomogeneous complete product is obtained, it is optionally cooled to atemperature below 60° C. and bottled.

Prior to further processing such as filling into capsules, it isfavorable to again stir the product to homogenize it, and if necessaryto heat it moderately, that is to a temperature of about 40° C. to 50°C. At a dilution ratio of 1:500 in water at a pH of 1.1 and atemperature of 37° C., the solubilizate exhibits an averaged turbidityof 1.0 FNU. The results of particle size analysis are summarized in thetable below.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 9.08 15.64292.7 615 Volume distribution 6.35 9.36 14.10 20.16

Exemplary Embodiment 4 Solubilizate of 5% Glavonoid (=1.8% Glabridin)/3%Curcumin/3.5% Xanthohumol

The following is used:

32 g 95% curcumin powder (=30.4 g of curcumin), 44 g Xantho-Flav Purepowder (=35.2 g of xanthohumol), 60 g Kaneka glavonoid (=1.8 gglabridin), 60 g 96% ethanol, neutral alcohol grade 1411U, 44 g 99.5%glycerol, 760 g  polysorbate 80.

Polysorbate 80 and glycerol are mixed together under stirring whilebeing heated to a temperature in the range from 48 to 52° C. tohomogenize the mixture adequately. Ethanol is incorporated into thepolysorbate-glycerol mixture while stirring intensely enough to form ahomogeneous solution, while the temperature is kept constant. Then,xanthohumol is incorporated into the solution of polysorbate, glycerol,and ethanol, while the temperature is raised to a value between 63 and67° C. under stirring intensely enough for the xanthohumol to combinehomogeneously with the prepared solution.

Subsequently, curcumin powder is incorporated into the xanthohumolsolubilizate, while the temperature is raised to a value in the rangebetween 78 and 82° C. As with the xanthohumol and also with theincorporation of glavonoid described below, stirring is performedintensely enough so that the newly added component of the solubilizatecombines homogeneously with the solubilized product in the preparedfluid. For the addition of glavonoid, the temperature is furtherincreased to a value in the range between 85 and 98° C.

The product is a solubilizate with co-micellated curcumin, xanthohumol,and glavonoid. It is allowed to cool to a maximum value of 45° C. whilestirring and is then bottled.

Exemplary Embodiment 5 Solubilizate of 1.5% Curcumin/2% Xanthohumol/3.5%Glavonoid (0.1% Glabridin)/1.2% Coenzyme Q₁₀

The following is used according to the formulations described above:

250 g 7% curcumin solubilizate, 250 g 9.2% xanthohumol solubilizate, 250g 15% glavonoid solubilizate, and 250 g 5% coenzyme Q10 solubilizate.

All four solubilizates can be heated to a temperature in the range from50° C. to 60° C. to lower viscosity and thus enhance flowability. Then,they are mixed together by stirring. Once a homogeneous complete productis obtained, it is optionally cooled to a temperature below 60° C. andbottled.

Prior to further processing such as filling into capsules, it isfavorable to again stir the product to homogenize it, and if necessaryto this end to heat it moderately, that is to a temperature of about 40°C. to 50° C.

The results of particle size analysis are summarized in the followingtable.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 8.22 10.8814.80 450 Volume distribution 7.78 10.06 13.10 16.39

Exemplary Embodiment 6 Solubilizate of 1.3% Curcumin/1.6% Xanthohumol/3%Glavonoid (0.09% Glabridin)/1% Coenzyme Q₁₀/2% α-Liponic Acid

The following is used according to the formulations described above:

200 g 7% curcumin solubilizate, 200 g 9.2% xanthohumol solubilizate, 200g 15% glavonoid solubilizate, 200 g 5% coenzyme Q₁₀ solubilizate, and200 g 10% α-lipoic acid solubilizate.

All five solubilizates can be heated to a temperature in the range from50° C. to 60° C. to lower viscosity and thus enhance flowability. Then,they are mixed together by stirring. Once a homogeneous complete productis obtained, it is optionally cooled to a temperature below 60° C. andbottled.

Prior to further processing such as filling into capsules, it isfavorable to again stir the product to homogenize it, and if necessaryto this end to heat it moderately, i.e. to a temperature of about 40° C.to 50° C.

The results of particle size analysis are summarized in the followingtable.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 8.85 11.6417.05 981 Volume distribution 7.16 9.41 12.36 15.43

Exemplary Embodiment 7 Solubilizate of 0.8% Curcumin/1.5% BoswellicAcid/1% Xanthohumol/1.8% Glavonoid (0.05% Glabridin)/0.18%Serrapeptase/1.2% Resveratrol/0.6% Coenzyme Q10/1.2% α-Liponic Acid

The following is used according to the formulations described above:

125 g 7% curcumin solubilizate, 125 g 12% Boswellia solubilizate, 125 g9.2% xanthohumol solubilizate, 125 g 15% glavonoid solubilizate, 125 g1.5% serrapeptase solubilizate, 125 g 10% resveratrol solubilizate, 125g 5% coenzyme Q₁₀ solubilizate, and 125 g 10% α-lipoic acidsolubilizate.

All eight solubilizates can be heated to a temperature in the range from50° C. to 60° C. to lower viscosity and thus enhance flowability. Then,they are mixed together by stirring. Once a homogeneous complete productis obtained, it is optionally cooled to a temperature below 60° C. andbottled.

Prior to further processing such as filling into capsules, it isfavorable to again stir the product to homogenize it, and if necessaryto this end to heat it moderately, i.e. to a temperature of about 40° C.to 50° C.

At a dilution ratio of 1:50 in water at a pH of 1.1 and a temperature of37° C., the solubilizate exhibits an averaged turbidity of 12.5 FNU.

The results of particle size analysis are summarized in the followingtable.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 8.09 10.9115.19 425 Volume distribution 6.90 9.26 12.37 15.71

Exemplary Embodiment 8 Solubilizate of 0.8% Curcumin/1.5% BoswellicAcids/1% Xanthohumol/1.8% Glavonoid (0.05% Glabridin)/0.18%Serrapeptase/1.2% Resveratrol/0.6% Coenzyme Q₁₀/1.2% α-Liponic Acid inDirect Preparation

The following is used:

9.375 g 95% curcumin powder, 19 g Boswellia Serrata extract (15.2boswellic acid), 12.5 g Xantho-Flav powder (at least 80% xanthohumol =10 g xanthohumol), 18.75 glavonoid (0.56 g glabridin), 1.875 gserrapeptase (37,500,000 U), 12.5 g resveratrol, 7.19 g coenzyme Q₁₀,12.5 g α-lipoic acid, 7.875 g water, 18.75 g ethanol, 12.5 g glycerol,9.375 g mixed tocopherol, 27.685 g MCT oil, 122.5 g polysorbate 20, and707.225 g polysorbate 80.

Polysorbate 80 is mixed with α-lipoic acid at a temperature in the rangebetween 18 and 22° C. while stirring intensely enough so that ahomogeneous mixture is created. Separately, the serrapeptase isdissolved in water in the same manner, at a temperature in the rangefrom 18 to 22° C. The further ingredients ethanol, xanthohumol,curcumin, Boswellia, glavonoid, resveratrol, coenzyme Q₁₀, mixedtocopherol, glycerol, MCT oil, and polysorbate 20 are slowly andsuccessively added to the mixture or solution of serrapeptase and waterunder constant stirring while the temperature is still in the rangebetween 18 and 22° C. Care is taken to ensure good mixing andhomogeneity of the product. If appropriate, pauses are taken between theaddition of a substance and the addition of the next substance. Theadding and stirring is performed slowly enough so that the particularingredient to be added is evenly incorporated into the preparation.

Next, both mixtures, i.e. the polysorbate 80 and the α-liponic acidpreparations, and the remaining mixture of all other ingredients aremixed together and further stirred at a temperature in the range from 18to 22° C. This creates a homogeneous pasty mass similar to a slurry,which is heated to a temperature in the range from 85° C. to 89° C. Theheating is performed under constant stirring and slowly enough so thatthe heated slurry always remains mixed as homogeneously as possible.After cooling to a temperature below 60° C., the product is bottled. Itis dark and viscous and is stored in the dark at temperatures of notmore than 25° C. Prior to further processing such as filling intocapsules, it is favorable to again stir the product to homogenize itand, if necessary to this end, to heat it moderately, i.e. to atemperature of about 40° C. to 50° C.

At a dilution ratio of 1:50 in water at a pH of 1.1 and a temperature of37° C., the solubilizate exhibits a turbidity of 1.0 FNU.

The results of the particle size analysis are summarized in thefollowing table.

d₁₀ (nm) d₅₀ (nm) d₉₀ (nm) d₉₉ (nm) Intensity distribution 8.66 11.0114.58 220.4 Volume distribution 7.96 9.87 12.59 15.52

Exemplary Embodiment 9 Solubilizate of 3% Curcumin/3.2% BoswellicAcid/1.6% Xanthohumol/1% CBD Oil

The following is used:

40.5 g 80% Boswellia serrata extract (32.4% boswellic acid), 31.5 g 95%curcumin powder (29.925 g of curcumin), 20.7 g Xantho-Flav powdercontaining at least 80% of xanthohumol (16.5 g xanthohumol), 54 g water,45 g ethanol, 315 g polysorbate 20, 483 g polysorbate 80, 10 g CBD oil:30% cannabidiol.

Cannabidiol (CBD) is a barely psychoactive cannabinoid derived from thefemale hemp Cannabis sativa or Cannabis indica. A non-THC-free CBD oilwas used,

1.-34. (canceled)
 35. A solubilizate, comprising: curcumin in a contentof 3 wt % to 7 wt %; at least one further active substance selected fromthe group consisting of xanthohumol, plant extracts, resin of afrankincense tree, licorice, cannabinoids, enzymes, serrapeptase,coenzyme Q₁₀, α-lipoic acid, and resveratrol; and at least oneemulsifier having an HLB value in a range below 18, namely polysorbate80 or polysorbate 20 or a mixture of polysorbate 20 and polysorbate 80,wherein the polysorbate content is at least 70 wt %.
 36. Thesolubilizate as claimed in claim 35, wherein the solubilizate containsup to 20 wt % of ethanol.
 37. The solubilizate as claimed in claim 35,wherein the solubilizate contains up to 25 wt % of glycerol.
 38. Thesolubilizate as claimed in claim 35, wherein the solubilizateadditionally contains up to 10 wt % of water.
 39. The solubilizate asclaimed in claim 35, wherein a diameter distribution of micelles in adilution of the solubilizate with distilled water in a ratio of 1:500under physiological conditions (pH 1.1 and 37° C.) is in a range fromd₁₀=6 nm to d₉₀=20 nm.
 40. The solubilizate as claimed in claim 35,wherein a turbidity of the solubilizate is less than 25 FNU, measured byscattered light measurement using infrared light according to thespecifications of the ISO 7027 standard at a dilution of thesolubilizate in a ratio of 1:50 or 1:500 in water under physiologicalconditions (pH 1.1 and 37° C.).
 41. A capsule filled with thesolubilizate as claimed in claim
 35. 42. A fluid, containing asolubilizate as claimed in claim 35, wherein the fluid is selected fromthe group consisting of foods, dietary supplements, beverages,cosmetics, and pharmaceutical products.
 43. The fluid of claim 42,wherein the fluid comprises an aqueous dilution of the solubilizate. 44.A method for treating and/or preventing diseases involving inflammation,cancer, Alzheimer's, Parkinson's, obesity, high cholesterol, elevatedblood sugar, diabetes, metabolic syndrome, and/or autoimmune diseases,multiple sclerosis (MS), for reducing visceral fat, for thermogenesis,for lowering cholesterol, for lowering LDL cholesterol and/or glucose inthe blood and/or triglycerides in the blood, for improving macularpigment density, for reducing oxidative stress and/or for reducing theaccumulation of fat in the hepatocytes, for treating and/or preventingfatty liver disease, Friedreich's ataxia, lysosomal diseases, Tay-Sachsdisease, arteriosclerosis, heart diseases, arthritis, comprising:administering, to a dietary supplement consumer or patient, thesolubilizate according to claim
 35. 45. The method of claim 44, whereinthe solubilizate is administered to the dietary supplement consumer orpatient in a dose of curcumin ranging from 0.5 mg/kg body weight to 1mg/kg body weight.
 46. The method of claim 44, wherein the solubilizateis administered to the dietary supplement consumer or patient in aBoswellia dose ranging from 1 mg/kg body weight to 2 mg/kg body weight.47. The method of claim 44, wherein the solubilizate is administered tothe dietary supplement consumer or patient in a xanthohumol dose rangingfrom 0.5 mg/kg body weight to 1 mg/kg body weight.
 48. A method forproducing the solubilizate as claimed in claim 35, comprising the stepsof: (a) providing polysorbate 80 and/or polysorbate 20 and/or a mixtureof polysorbate 20 and polysorbate 80; (b) adding at least one furtheractive substance; and (c) adding curcumin powder, wherein step (a)comprises heating to a temperature in a range from 40° C. to 62° C., andwherein step (b) comprises keeping the temperature unchanged compared tostep a), or heating to a temperature in a range from 60° C. to 75° C.,and wherein step (c) comprises heating to a temperature in a range from82° C. to 97° C.
 49. The method of claim 48, wherein prior to step (b),a step (b1) is performed, comprising adding water at a temperature in arange from 40° C. to 62° C.
 50. The method of claim 49, wherein step(b1) further comprises adding ethanol at a temperature in a range from40° C. to 62° C.
 51. A method for producing a solubilizate as claimed inclaim 35, comprising the steps of: (a) preparing a first preparation byproviding polysorbate 80 and/or polysorbate 20 and/or a mixture ofpolysorbate 20 and polysorbate 80 and at least a first active substance;(b) preparing a second preparation by providing water and at least onefurther active substance; (c) adding at least one additional activesubstance, and adding curcumin powder to the second preparation fromstep (b); (d) combining the first preparation from step (a) and thesecond preparation from step c), wherein the temperature is in a rangefrom 18° C. to 22° C. during the execution of steps (a) through (d); and(e) heating to a temperature in a range from 80° C. to 97° C.
 52. Themethod of claim 51, wherein step (c) further comprises adding ethanoland/or glycerol and/or MCT oil and/or polysorbate 20 and/or polysorbate80 and/or a mixture of polysorbate 20 and polysorbate
 80. 53. A methodfor producing a solubilizate as claimed in claim 35, comprising thesteps of (a) providing polysorbate 80 and/or polysorbate 20 and/or amixture of polysorbate 20 and polysorbate 80 and of glycerol and ofethanol; (b) adding at least one further active substance, wherein step(a) comprises heating to a temperature in a range from 40° C. to 62° C.,and wherein step (b) comprises heating to a temperature in a range from60° C. to 75° C.; (c) adding curcumin powder at a temperature in a rangefrom 70° C. to 92° C.; and (d) adding glavonoid under heating to atemperature in a range from 80° C. to 97° C.
 54. A method for producingthe solubilizate as claimed in claim 35, by mixing a curcuminsolubilizate and a solubilizate of at least one further activesubstance.
 55. A method for producing a xanthohumol solubilizate, foruse in a method according to claim 54, comprising the steps of (a)providing an ethanolic extract of hard resins from hops; and (b) addingpolysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20and polysorbate 80, wherein step (b) comprises heating to a temperaturein a range from 80° C. to 95° C.
 56. The method of claim 55, whereinprior to step (b), a step (b1) is performed, comprising dissolving theethanolic extract of the hard resins from hops, in ethanol under heatingto a temperature in a range from 40° C. to 62° C.
 57. A method forpreparing an enzyme solubilizate, for use in a method according to claim54, comprising the steps of (a) providing water and enzyme; (b) addingMCT oil; and (c) adding polysorbate 80 and/or polysorbate 20 and/or amixture of polysorbate 20 and polysorbate 80, wherein the temperature isin a range from 18° C. to 22° C. while performing step (a) and whereinstep (b) comprises heating to a temperature in a range from 77° C. to93° C.